Malaria causes 500 million clinical cases and > 1 million deaths annually, is responsible for a loss of > 1.3% of GDP in Africa annually, and is a serious concern for travelers and the military. A vaccine would be the ideal preventive and is perhaps the only intervention that could facilitate eradication of the disease. When attenuated P. falciparum sporozoites (PfSPZ) are administered by bite of infected mosquitoes, > 90% of human volunteers are protected against experimental Pf challenge and protection lasts at least 10 months. Sanaria's goal is to develop and commercialize an attenuated PfSPZ vaccine that produces > 90% protective efficacy, for 3 markets with a potential for > $1 billion annual revenues; 1) Travelers from the developed world, 2) Infants and young children in the developing world, and 3) Adolescent girls in the developing world. In a separate Phase II SBIR and with other funds Sanaria has developed and manufactured under current Good Manufacturing Practices (cGMPs) the PfSPZ vaccine using the cryopreservation and irradiation methods developed in our Phase I SBIR. The vaccine met all release specifications, is now in pre-clinical IND-enabling toxicology studies, and is expected to enter clinical trials in the U.S. by the last quarter of 2008. The goals of this proposal are to facilitate rapid testing in Africa by translating the successful cryopreservation protocol developed in Phase I, into a comprehensive system to support clinical trials in Africa, and to facilitate scale up, large scale Phase 3 clinical trials, and vaccine launch by establishing the capacity for scaled-up vaccine production in the area of cryopreservation. This proposal has 2 major components, 1) Work designed to provide the foundation for vaccine distribution in expanded Phase 2 and Phase 3 clinical trials, and during commercialization (Specific Aims 1-3), and 2) Work designed to scale up vaccine production for Phase 3 trials and commercialization (Specific Aims 4-6). The Specific Aims are to: 1. Develop methods for storage, inventory control and recovery of vaccine batches for shipment and use in clinical trials; 2. Establish and validate a below -140oC cold chain for all vaccine movements and all transit points between site of vaccine manufacture and testing centers in the U.S., and expansion of methods and infrastructure for vaccine delivery to Africa; 3. Establish in-clinic protocols for thawing and diluting cryopreserved vaccine and preparation for injection, and development and validation of vaccine point-of-administration protocols for use under conditions in Africa; 4. Extend vaccine viability and potency during cryopreservation steps; 5. Create initial daily cryopreserved vaccine production sub-lots, which are then combined into single large vaccine lots, and 6. Automate the protocol for cryopreserving large vaccine lots. These Aims will include establishing quality GMP/GCP control procedures as units of vaccine move to the clinic. There is an urgent need to extensively test the PfSPZ vaccine in Africa as soon as possible. Achievement of the Aims of this proposal will facilitate this testing and decrease time and cost to registration of the PfSPZ vaccine, thereby facilitating delivery of the vaccine to the African children, who need it most. PUBLIC HEALTH RELEVANCE: Malaria causes 500 million clinical cases and 1-3 million deaths annually, is responsible for >1% loss of GDP in Africa annually and is a serious concern for travelers and military personnel. Sanaria's goal is to develop and commercialize a >90% protective malaria vaccine for three primary markets with a potential for >$1 billion annual revenues: 1) Travelers from the developed world to malaria endemic areas. 2) Infants and young children in the developing world. 3) Adolescent girls in the developing world. Success in this project will significantly increase the capacity to produce larger vaccine lots at the same time decreasing cost of production; will establish routes for delivery of the vaccine to clinical trials sites in the USA and, importantly, to the populations at need in Africa. [unreadable] [unreadable] [unreadable]